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. 2020 May 25;12(2):107–127. doi: 10.1007/s13238-020-00723-9

Figure 3.

Figure 3

Loss of 4.1N promotes anoikis resistance of EOC cells in vitro. (A) Representative images of flow cytometry results with 7AAD–Annexin V-AF647 staining, A2780 WT, A2780 OE, SKOV3 WT and SKOV3 KO cells in suspension (left); results means ± SD from three independent experiments (right). (B) Representative images of A2780 WT, A2780 OE, SKOV3 WT and SKOV3 KO cells subjected to JC-1 staining assays (top); The ratio of red/green fluorescence indensity of JC-1 assay was representation on graphical (bottom). Scale bar, 50 µm. (C) Proliferation rate of anoikis resistant A2780 WT, A2780 OE, SKOV3 WT and SKOV3 KO cells was analyzed by comparing cell densities after 24 h incubation. (D) Cell viability was first evaluated using the trypan blue assay. (E) Effects of 4.1N on apoptosis-related mRNA assayed by qRT-PCR and results means ± SD from three independent experiments. β-Actin was used as internal control. (F) Effects of 4.1N on apoptosis-related proteins expression was assayed by Western blot. β-Actin was loading control. *P < 0.05, **P < 0.01, ***P < 0.001